This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Individuals with Familial Hypertriglyceridemia (FHTG) have elevated levels of triglycerides and normal total cholesterol. FHTG affects about 1% of the US population and is one of the most common genetic lipid disorders in patients with coronary artery disease. FHTG is thought to segregate as an autosomal dominant disorder but the single segregation analysis has not been conclusively replicated. The long-term objective of this proposal is to identify gene(s) that predispose individuals to FHTG using familial-based linkage analysis and determine how elevated levels of serum triglycerides and lipids predispose these individuals to atherosclerosis. We plan to accomplish the objective by pursuing the following two specific aims. 1. Identify Families with FHTG. We will identify probands through the statewide CARDIAC program and through a network of health clinics. 2. Identify FHTG susceptibility loci by linkage analysis on a genome-wide set of markers and on a small set of markers previously identified FHTG candidate loci. We will use linkage analysis methods to determine which loci contribute to FHTG in patients from the WV population and test for linkage and association with single nucleotide polymorphisms (SNPs) that are known to map to the regions of interest. Individuals with Familial Hypertriglyceridemia (FHTG) have elevated levels of triglycerides and normal total cholesterol. FHTG affects about 1% of the US population and is one of the most common genetic lipid disorders in patients with coronary artery disease. FHTG is thought to segregate as an autosomal dominant disorder but the single segregation analysis has not been conclusively replicated. The long-term objective of this proposal is to identify gene(s) that predispose individuals to FHTG using familial-based linkage analysis and determine how elevated levels of serum triglycerides and lipids predispose these individuals to atherosclerosis. We plan to accomplish the objective by pursuing the following two specific aims. 1. Identify Families with FHTG. We will identify probands through the statewide CARDIAC program and through a network of health clinics. 2. Identify FHTG susceptibility loci by linkage analysis on a genome-wide set of markers and on a small set of markers previously identified FHTG candidate loci. We will use linkage analysis methods to determine which loci contribute to FHTG in patients from the WV population and test for linkage and association with single nucleotide polymorphisms (SNPs) that are known to map to the regions of interest. The proposed work is innovative because we have access to patients in West Virginia who have not been studied. Our findings will be significant because they will further our undrstanding of the pathogenesis of vascular disease and because these susceptibility genes represent new targets for preventative and therapeutic interventions. Modification to Specific Aims: Specific Aims for this project were modified for the 2007-2008 funding cycle and approved by the WVINBRE External Advisory Committee (EAC) and the NIH. These modifications were necessary because of the inability to recruit patients into the Genetic Basis of Familial Hypertriglyceridemia (FHTG) study. Because of the change in specific aims in year four, this project has been re-titled "Effect of Statins on Inflammatory Response in Human Aortic Endothelial Cells (HAEC)". Dr. Donald Primerano, ACoRN director, has recommended termination of the FHTG project in favor of continuation of work on the modified specific aims. This recommendation will be considered by the Steering Committee and EAC in the near future. Dr. Kreisberg will continue to perform TaqMan Allelic discrimination assays in year five as needed for the Genetic Basis of Familial Combined Hyperlipidemia in addition to completion of the modified specific aims. Effect of Statins on Inflammatory Response in Human Aortic Endothelial Cells (HAEC) Modified Specific Aims: 1. Inhibition of the immune response by statins: We will determine whether statins inhibit the induction of interleukin 8 (Il-8) and VCAM by C-reactive protein (CRP), TNF, and oxidized-LDL in human aortic endothelial cells (HAEC). Cells will be treated with the CRP, TNFalpha, and oxidized LDL in the presence or absence of atorvastatin. Il-8 and VCAM mRNA and protein levels will be measured after treatment. To determine whether a statin effect is due to its action on HMG-CoA reductase, cells will be treated with inflammatory agents in the presence of atorvastatin and mevalonate. 2. Mechanisms by which statins protect HAEC from injury: We will determine the role of the Ras family of small GTPases in this response to injury. We will determine the effect of the inflammatory mediators on Ras, RhoA, and Rac activities, protein levels and gene expression.